Nascent membrane and presecretory proteins synthesized in Escherichia coli associate with signal recognition particle and trigger factor
Nascent membrane and presecretory proteins synthesized in Escherichia coli associate with signal recognition particle and trigger factor
复制标题
DOI：
10.1046/j.1365-2958.1997.4431808.x
复制
发表时间：
1997-07-01
影响因子：
3.6
通讯作者：
Luirink, J
中科院分区：
文献类型：
--
作者：
Valent, QA;deGier, JWL;Luirink, J
大肠杆菌信号识别颗粒（SRP）和触发因子是与在异源体外翻译系统中产生的分泌前和膜蛋白的短新生多肽相互作用的细胞质因子。在这项研究中，我们使用一个E。大肠杆菌体外翻译系统与双功能交联剂的组合，以研究这些相互作用在同源环境中更详细。使用这种方法，SRP与暴露特别疏水的靶向信号的新生多肽的直接相互作用被证明，这表明内膜蛋白是E. coli SRP。有证据表明，过度生产的蛋白质，暴露疏水多肽伸展，tillage SRP。此外，触发因子与不同长度和性质的新生多肽有效交联，一些短至57个氨基酸残基，表明其位于E.大肠杆菌核糖体。
The Escherichia coli signal recognition particle (SRP) and trigger factor are cytoplasmic factors that interact with short nascent polypeptides of presecretory and membrane proteins produced in a heterologous in vitro translation system. In this study, we use an E. coli in vitro translation system in combination with bifunctional cross-linking reagents to investigate these interactions in more detail in a homologous environment. Using this approach, the direct interaction of SRP with nascent polypeptides that expose particularly hydrophobic targeting signals is demonstrated, suggesting that inner membrane proteins are the primary physiological substrate of the E. coli SRP. Evidence is presented that the overproduction of proteins that expose hydrophobic polypeptide stretches, titrates SRP. In addition, trigger factor is efficiently cross-linked to nascent polypeptides of different length and nature, some as short as 57 amino acid residues, indicating that it is positioned near the nascent chain exit site on the E. coli ribosome.